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Organic components in effluents from mills for processing waste paper, prior to and after treatment, were studied by gas chromatography. Fatty acids, esters, phenols and alcohols at concentrations from 0.01 to 0.5 mg/l were determined.

The combination of affinity chromatography and immunoassay provides a novel method for the quantitation of steroids in biological fluids. Anti-digoxin and anti-oestriol-16α-glucuronide antibodies were immobilised to cellulose strips, and used for ascending paper chromatography. Labelled digoxin and oestriol-16α-glucuronide were selectively retarded on strips containing the appropriate immobilised antibodies. The degree of retardation was quantitatively affected by the addition of non-labelled steroid to the sample. Calibration curves could, therefore, be constructed. The sensitivity of the assay system can be altered by changing the concentration of immobilised antibody on the cellulose strip. Calibration curves over the range 50–800 pg for digoxin and 50–250 pg for oestriol-16α-glucuronide are presented.

Twenty-five steroidal glucosiduronic esters were chromatographed on paper in each of five straight-phase systems and five reversed-phase systems. The resolving characteristics of these systems were compared by expressing the retention values in terms of R M and correlating the data by use of linear regression equations. The resolving characteristics of the straight-phase systems as a group are very similar, and those of the reversed-phase systems are somewhat less similar; the various systems differ considerably in polarity. The resolving properties of a straight-phase system and a reversed-phase system which contain the same liquid ion exchanger are markedly different, a circumstance which increases the probability that a pair of compounds which was not separated in a straight-phase system will be resolved in the reversed-phase system which contains the same liquid ion exchanger. The migration of the esters in the foregoing systems was compared with the migration of the corresponding glucosiduronic acids (a previous study) in the same solvent systems. If a particular pair of conjugates is not separable as carboxylic acids in a specific straight-phase system, there is a relatively good chance that they will be separable as methyl esters in the same system.

Novel chromatographic and derivatization techniques have been developed to investigate the levels and kinetics of 3-phosphoglyceric acid, phosphohydroxypyruvate and phosphoserine in plants. These techniques use derivatization reactions of the phosphorylated compounds with o -phenylenediamine and isonicotinic acid hydrazide, in combination with paper chromatography. Unexpected reactions of α-hydroxy acids have been encountered and circumvented.

R F values of 41 cobalt(III) complexes were determined by partition paper chromatography using 15 solvent systems. It was established that cis -NO 2 isomers always exhibit higher R F values than the corresponding trans -isomers, even when a complex has several cis - and trans -isomers. It has been found that an increase in the chelate ring size gives rise to higher R F values and that there is a linear dependence between the R M values observed and the number of five-membered rings that are substituted by the corresponding six-membered rings.

The electrophoretic mobility of an α-hydroxycarboxylic acid in electrolytes containing boric acid is very dependent on the degree of substitution at the α-carbon atom. A study of eighteen α-hydroxy acids shows clearly that the “ gem -dialkyl” effect is operating to favour the formation of the strongly acidic boric acid complexes from the more densely substituted acids, the mobilities of whcih are thus more or less greatly enhanced. In general, the complexes from best at pH 2, but some are stable at higher levels. Of several that exist at pH 4.6, only the complexes of viridifloric, trachelanthic and lasiocarpic acids also survive paper electrophoresis at pH 9.2. These acids form 1:1 instead of the usual 1:2 complexes and they are further distinguished from the others by an unusually high degree of crowding at their α-carbons, and by containing a β- as well as the α-hydroxyl group.

The action of HClO 4 on rhodium hydroxide was studied with paper electrophoresis and ion exchange chromatography. The existence of chloro-complexes due to precipitation of co-ordinated Cl in the preparation of Rh(OH) 3 , was demonstrated. These can be decomposed by repeated precipitation or heating with AgNO 3 .

The RF′ values of a number of isomeric monosubstituted phenols have been determined for three solvent systems. This is a continuation of our previous data. The results confirm the earlier work showing that most of the phenols have significant variations in RF′ value between these three solvents. Some limited correlations between RF′ values and the molecular weight of the side chain were found for members of homologous series. The use of ceric ammonium nitrate as a spot locating agent for phenols was found to have certain limitations, particularly for ortho substituted phenols. In general, however, this reagent is quite suitable for most phenols.

A chromatographic separation on glass fiber paper of mixed glycerides of acetic, butyric, stearic, palmitic, and oleic acids in solvents consisting of 2% and 5% ether in isooctane has been described. In general, glycerides containing fatty acids of increasing chain length tend to move further up the chromatograms than those of shorter chain length.

A method was developed, combining reaction paper chromatography with electrolytic reduction, for the determination of the presence of -COOH groups and possibly their number by comparing the RF values before and after reaction. Moreover, in the solvent system used (n-propanol-ammonia), it is possible to assess the presence of other functional groups from the shift in the RF values.

Summary Some of the compounds of Sn(II) with platinum metals may be separated by adsorption chromatography on paper. Their behaviour in SnCl 2 -HCl and SnBr 2 HBr was noted.

A simple technique for measuring diffusion rates on glass fibre paper is described. This technique was shown to be suitable for the study of polyacid formation and complexing in solution.

The adsorption paper chromatography of inorganic anions in acetate buffers was studied and the possible adsorption mechanism discussed. Some analytical separations were recorded.

Rapid reliable methods have been developed for the separation of mixtures of organic acids using one-dimensional and two-dimensional techniques of paper chromatography. New solvent mixtures are proposed which include acidic, basic and buffered types. These methods, which have been proved by application to some eighty well-known organic acids, offer considerable advantages over existing techniques in their ease of operation and ready reproducibility of results.

Operating variables in the use of recording ratemeters for scanning radioactive paper chromatographic strips are discussed. Bes sensitivity, and least statistical error in estimation of peak area, is obtained with a wide slit. However, wide slits give poor fidelity, that is, the shape of the record is not an accurate representation of the shape of the actual peak. Short time constnts give a record showing undesirable fluctuations due to statistical count rate variations, while long time constants give poor fidelity. Slow strip speed is required for strips of low activity. Optimal selection of operating conditions is considered from a quantitative standpoint.

Mean R F and R M values have been obtained for a number of steroids and dyes in Bush aqueous methanol systems. Scrutiny of the Δ R M values verifies the stability of the condition achieved.

Summary The chromatographic behaviour of metal ions on acetylated paper with butanol-HCl was investigated. The lower valencies of polonium may be differentiated in this system.

The separation of cis- and trans-dihalogenodiammineplatinum (II) complexes was effected by paper chromatography using neutral fast flowing solvents such as water at 5° or acetone—water (9:1). Practically all mixtures of cis- and trans- as well as of dichloro-, dibromo-, diiodo as well as chloroiodo- and chlorobromo-complexes could be separated.

The use of ethylene glycol as the stationary phase for the separation of corticosteroids and of 17-ketosteroids by paper chromatography is proposed. Some characteristics of systems with ethylene glycol in the stationary phase are described.

The authors studied the use of chloranilic acid as a reagent in the paper chromatography of some of the heavier metals. Sensitivity was found to vary from 0.01 μg for ferric iron, to 5.0 μg for mercury (II) and zinc. It was found that iron gave tailing spots with most of the solvent systems, which interfered with the separation of the other cations. This interference was avoided by using acetone—acetic acid, which displaces iron to the solvent front. Efficient two-dimensional separations were obtained by carrying out a second run with ethanol—acetic acid.

The object of this work was to find a rapid and precise method for the separation of the principal monosaccharide constituents of the carbohydrate fraction of wood: glucose, galactose, mannose, arabinose and xylose. Discarding the imprecise methods of classical analysis, we have considered the possibility of the application of paper chromatographic methods. None of the eluents suggested in the literature gives a satisfactory solution to this problem. We have studied the use of a solvent composed of a mixture of ethyl acetate, pyridine and water in the proportions 3.6:1:1.15; we have found that this eluent gives a complete separation of the sugars and allows both qualitative and quantitative determinations.

The supernumerary (third) spots which separate when binary mixtures of copper(II) complexes of histidine and some other amino acids are subjected to paper electrophoresis were re-examined. It is concluded that, in the early stages of electrophoresis, the spots consist of mixed-ligand complexes composed of copper(II), histidine and other amino acid and that, as electrophoresis proceeds, the complexes undergo tow different reactions, simultaneously. A portion of them re-equilibrates to form homogeneous copper(II) complexes of each of their constituent amino acids, while the remainder decomposes to the relatively stable 1:1 histidine—copper(II) complex and the other amino acid which is liberated as the free, uncomplexed ion. This interpretation is based on electrophoresis in mildly alkaline electrolytes of mixtures containing copper(II) histidnate and the copper(II) complex of either an acidic or a basic amino acid.

Methods are given for the separation and analysis of(i) the methylated sugars obtained from the methylated polysaccharide, and (ii) the glycosides obtained by a SMITH degradation of the mannan. Analysis of a mixture of isomeric methylated sugars was facilitated by successive borohydride reduction, periodate oxidation and borohydride reduction. In this way isomeric C6 sugars were selectively degraded into C6, C5, C4, C3 and C2 alditols which were readily separated by vapour-phase chromatography (v.p.c.). Details are given for the separation of tri-, di-, and mono-O-methyl-d-mannoses. SMITH degradation of the mannan gave a series of glycosides of which i-O- (α- and β-d-mannopyranosyl)-glycerol, i-O-(α- and β-d-lyxofuranosyl)-erythritol and 2-O-(d-lyxofuranosyl)-d-erythritol were separable by paper chromatography. Other glycosides separated by v.p.c. of their trimethylsilyl derivatives were 2-O-(α-d-mannopyranosyl)-d-erythritol, the β-anomer (p-nitrobenzoate, m.p. 235-238°); i-O-(β-d-mannopyranosyl)-erythritol (p-nitrobenzoate, m.p. 239–243°) and the αanomer. The mannan was shown to contain d-mannofuranose and d-mannopyranose residues, to be highly branched and to contain mainly (i → 4) and (i → 6) linkages.

Four different chromatographic solvents containing borates of organic bases are described. Two of them permit the separation of sugar nucleotides differing only in the sugar moiety and give also a good resolution for some sugar 1-phosphates. The other two systems permit, in a single run, the separation of a group of sugars which is usually difficult to achieve.

Selected organic compounds commonly occurring in plant and animal tissues have been chromatographed on glass fiber, paraffin-coated and untreated cellulose paper in a number of different solvent systems. The RF values of these compounds represent new additions to the literature in the solvent systems studied.

Solubilized vat dyes wre separated by paper chromatography, using a methanolic solution of ammonia as solvent system. A system containing pyridine, isoamyl alcohol and ammonia, as well as a mixture of methanol and acetic acid were also used. Both descending and centrifugal chromatography were employed. The relation between dye constitution and chromatographic behaviour has been examined.

The low-voltage and high-voltage paper electrophoretic behaviour of iodide, iodate, periodate, tellurite and tellurate ions has been studied in sodium hydroxide and sodium chloride solutions used as electrolyte. Iodide, iodate and periodate can be very well separated in 0.01–1.0 N NaOH or in sodium chloride solutions of concentration less than 0.5 N. Simultaneous separation of iodide, iodate, periodate, tellurite and tellurate could not be realised. One could, however, obtain the following separations: tellurate, tellurite, (iodate + periodate), and iodide in NaCl by low-voltage electrophoresis, periodate, tellurite, iodate and iodide by high-voltage electrophoresis in sodium chloride (< 0.5 N).

Summary Six solvent systems, two of them of “reversed phase” type, are recommended for the paper chromatographic separation of narcotics. A chromatographic pattern obtained in these systems will facilitate the identification process.

An acidic and a basic solvent containing the same proportions of propanol and water have been used for the study of about 100 organic acids by paper chromatography. The acid and the base were solutions fo the gases sulphur dioxide or ammonia in water and therefore were readily removed after chromatography. Sulphurous acid has the advantage of being a stronger acid than those normally used, e.g. formic and acetic acid. The R M values of the organic acids in the two solvents can be arranged into clear-cut groups dependent on the number of carboxyl and other substituted groups present, in such a way that the number of carboxyl groups in an unknown acid can be confidently predicted. An indication can also be obtained of other groups such as alkyl, aryl, amino, hydroxy, bromo etc. which may be present, and the manner in which they are arranged. The results include nine homologous series whose members differ from one another only in the addition of -CH 2 - to the molecule.

N-(2-Hydroxyethyl)iminodiacetic acid (HIDA) is a very suitable complexing agent for paper electrophoretic separations of inorganic ions. It is fairly soluble even in an acidified aqueous solution. Its complexes migrate as well defined zones with different mobilities, depending on their stability. The electrophoretic behaviour of 64 inorganic ions in solutions of HIDA as a function of pH value was studied (electrophoretic mobility curves). Separation at pH 2–3 yields the best results for many metal ions; some further separations succeed in an alkaline solution (pH 9–10). Examples of separations of a number of model mixtures are given.

To supplement extensive information on peptides and amino acids, data were obtained on the paper-chromatographic behavior of 88 synthetic peptides and 29 amino acids in 15 solvent systems. Theoretical R F values were calculated for 46 peptides in 9 solvent system and were found to be in good agreement with the observed values.

The R F values are given of about 40 inorganic anions in a number of isopropanol-1.5 N ammonia mixtures, as well as on anion exchange resin paper with N KCl or N KNO 3 as solvent. These data together with the electrophoretic mobilities in o.r N NaOH are used in a scheme of identification based on the principle of “ R F spectra”.

Previous experimental and theoretical work on quantitative paper partition chromatography by spot-area and spot-length is reviewed. An asymmetric diffusion model for chromatography, where diffusion in the direction of developer flow and perpendicular to it are unequal by virtue of the kinetics of partition, is applied to derive the relationships that spot-area is directly proportional to log spot-content, and spot-length is directly proportional to square-root of log spot-content. Relations for the increase of spot-area and apot-length with time are given.

It has been shown by these examples that the technique of paper electrophoresis, using inexpensive equipment which can be put together in almost any laboratory, is capable o f separating many of the radiactive ionic species formed in typical S zilard -C chalmers reactions, and is also useful in assisting in their identification.

Paper chromatography has proved to be suitable for the separation of both Degranol and Mannitol-Myleran. Experiments made in this connection showed that Degranol was more stable than the other agent.

A detailed study has been made of the sorption of copper (II) ions by Whatman 41 filter paper. With low concentrations of copper ion, adsorption of the cation can be attributed to ion exchange, at higher values ingestion of the salt into the fibres occurs. The observed relationship between the extent of adsorption and the exchange capacity is explained in terms of a hyphotesis based on the Donnan membrane theory.

Resume Ion pair formation between cobalt(III) complexes and organic anions and between anionic complex cyanides and polyvalent metal ions was studied by paper electrophoresis. It was shown that complexes of the type [CoA 6 ] 3+ can move anionically (for example in trichloroacetate) and that complexes of the type [M(CN) 6 ] 2−or3− can move cationally (for example in aluminum or zirconium chloride).

A method is described for the determination of the biologically active components of the organophosphorus pesticide VUAgT-3 in technical products and commercial formulations. The procedure consists of chromatographic separation on formamide impregnated papers with n-hexane and toluene as solvent, and colorimetric determination of phosphorus in eluates of evndetected spots.

An investigation was carried out of the chromatographic behaviour of some alkyl derivatives of phenol, pyrocatechol, hydroquinone, and 4,4′-isopropylidene-bisphenol using papers impregnated with formamide or monomethylformamide. The R F values of the compounds were related to a homologous series of standards; pyrocatechol and 4-alkylpyrocatechols containing C 1 —C 12 alkyls were used for this purpose and were assigned to R C values. The Δ R C contributions of the individual groups were established, and their additivity was verified.

A study was made of the determination of food preservatives by direct extraction and isolation on chromatographic paper. Recovery experiments from fruit juice (sorbic and benzoic acids) and from fish (benzoic acid and p-hydroxybenzoates) were carried out.

Summary The separation of Ca-Sr-Ba was achieved by paper chromatography using concentrated nitrate solutions with propanol as solvent. Ba yields double spots with some organic solvents. The method enables a separation of Ba and Ra in trace amounts and is useful for the radiochemical purification of 140 Ba and 228 Ra sources.